1. Date of download: 6/21/2016 Copyright © The American College of Cardiology. All rights reserved. From: Left ventricular diastolic dysfunction during demand ischemia: rigor underlies increased stiffness without calcium-mediated tension. Amelioration by glycolytic substrate J Am Coll Cardiol. 2001;37(8):2144-2153. doi:10.1016/S0735-1097(01)01282-7 Calcium during demand ischemia. Ischemia (decreasing coronary perfusion pressure [CPP] to 20 mm Hg) reduced left ventricular systolic pressure (LVSP) to 70 mm Hg and left ventricular end-diastolic pressure (LVEDP) to 13 mm Hg. Tachycardia increased isovolumic LVEDP to 20 mm Hg, indicating increased diastolic chamber stiffness. A 14 mmol/L calcium (14 mM Ca ++ ) imposed after tachycardia during sustained ischemic diastolic dysfunction increased LVSP from 60 to 110 mm Hg, indicating increased intracellular calcium concentration. However, LVEDP did not simultaneously increase (but in this example decreased from 20 to 18 mm Hg), suggesting that during ischemic diastolic dysfunction, calcium resequestration mechanisms remained intact and that the upward shift in LVEDP during tachycardia was not calcium-driven. Left ventricular systolic pressure recovered subsequent to infusion. Figure Legend:

2. Date of download: 6/21/2016 Copyright © The American College of Cardiology. All rights reserved. From: Left ventricular diastolic dysfunction during demand ischemia: rigor underlies increased stiffness without calcium-mediated tension. Amelioration by glycolytic substrate J Am Coll Cardiol. 2001;37(8):2144-2153. doi:10.1016/S0735-1097(01)01282-7 Ammonium chloride (NH 4 Cl) during demand ischemia. Ischemia (decreasing coronary perfusion pressure [CPP] to 20 mm Hg) reduced left ventricular systolic pressure (LVSP) to 60 mm Hg and left ventricular end-diastolic pressure (LVEDP) to 18 mm Hg. Tachycardia increased LVEDP to 25 mm Hg, indicating increased diastolic chamber stiffness. 50 mmol/L ammonium chloride (50 mM NH 4 Cl) imposed after tachycardia during sustained ischemic diastolic dysfunction initially increased LVSP from 63 to 80 mm Hg, then reduced it to 56 mm Hg, indicating a positive, followed by a negative, inotropic effect, consistent with its known effects on intracellular pH and, hence, myofilament calcium-sensitivity. Elevated LVEDP, however, remained unchanged, implying that it was not increased by a calcium-activated mechanism. Figure Legend:

3. Date of download: 6/21/2016 Copyright © The American College of Cardiology. All rights reserved. From: Left ventricular diastolic dysfunction during demand ischemia: rigor underlies increased stiffness without calcium-mediated tension. Amelioration by glycolytic substrate J Am Coll Cardiol. 2001;37(8):2144-2153. doi:10.1016/S0735-1097(01)01282-7 Egtazic acid (EGTA) during demand ischemia. Ischemia (decreasing coronary perfusion pressure [CPP] to 20 mm Hg) reduced left ventricular systolic pressure (LVSP) to 68 mm Hg and left ventricular end-diastolic pressure (LVEDP) to 15 mm Hg. Tachycardia increased LVEDP to 21 mm Hg, indicating increased diastolic chamber stiffness. An 8 mmol/L EGTA (8 mM) imposed after tachycardia during sustained ischemic diastolic dysfunction reduced LVSP from 70 to 48 mm Hg, indicating reduced intracellular calcium concentration but did not decrease elevated LVEDP, suggesting that this was not calcium-driven. Contractile function recovered after termination of EGTA. Figure Legend:

4. Date of download: 6/21/2016 Copyright © The American College of Cardiology. All rights reserved. From: Left ventricular diastolic dysfunction during demand ischemia: rigor underlies increased stiffness without calcium-mediated tension. Amelioration by glycolytic substrate J Am Coll Cardiol. 2001;37(8):2144-2153. doi:10.1016/S0735-1097(01)01282-7 Butane-dione-monoxime (BDM) during demand ischemia. Ischemia (decreasing coronary perfusion pressure [CPP] to 20 mm Hg) reduced left ventricular systolic pressure (LVSP) to 62 mm Hg and left ventricular end-diastolic pressure (LVEDP) to 18 mm Hg. Tachycardia increased LVEDP to 28 mm Hg, indicating increased diastolic chamber stiffness. A 5 mmol/L BDM (5 mM) infusion imposed after tachycardia during sustained ischemic diastolic dysfunction progressively decreased LVSP from 62 to 43 mm Hg, indicating inhibition of calcium-activated myofilament cross-bridge cycling. However, elevated LVEDP was not simultaneously decreased, suggesting that it was not calcium-driven. Contractile function recovered after termination of infusion. Figure Legend:

5. Date of download: 6/21/2016 Copyright © The American College of Cardiology. All rights reserved. From: Left ventricular diastolic dysfunction during demand ischemia: rigor underlies increased stiffness without calcium-mediated tension. Amelioration by glycolytic substrate J Am Coll Cardiol. 2001;37(8):2144-2153. doi:10.1016/S0735-1097(01)01282-7 Increased glycolytic substrate during demand ischemia. Ischemia (decreasing coronary perfusion pressure [CPP] to 20 mm Hg) reduced left ventricular systolic pressure (LVSP) to 55 mm Hg and left ventricular end-diastolic pressure (LVEDP) to 16 mm Hg. Tachycardia increased LVEDP to 24 mm Hg, indicating increased diastolic chamber stiffness. High 25 mmol/L glucose (25 mM) and insulin (400 μU/ml) infusion gradually increased LVSP, and concomitantly decreased LVEDP from 24 to 18 mm Hg (i.e., reversed the increase in LV diastolic chamber stiffness produced by demand ischemia), toward pre-tachycardia values. Figure Legend:

6. Date of download: 6/21/2016 Copyright © The American College of Cardiology. All rights reserved. From: Left ventricular diastolic dysfunction during demand ischemia: rigor underlies increased stiffness without calcium-mediated tension. Amelioration by glycolytic substrate J Am Coll Cardiol. 2001;37(8):2144-2153. doi:10.1016/S0735-1097(01)01282-7 Summary of hemodynamic effects of increased glycolytic substrate during demand ischemia, contrasted with equiosmotic mannitol. Groups were similar before intervention. Tachycardia (7 Hz) imposed during ischemia increased diastolic stiffness (isovolumic left ventricular end-diastolic pressure [LVEDP]). Glucose 25 mmol/L and insulin 400 μU/ml (G + I) progressively reduced increased diastolic stiffness and enhanced contractile function (left ventricular developed pressure [LV Dev P]). Mannitol had no effect. Figure Legend: